GSK3B Overexpression Alleviates Posttraumatic Osteoarthritis in Mice by Promoting DNMT1-Mediated Hypermethylation of NR4A3 Promoter

Figure 4

NR4A3 promoter hypermethylation alleviates IL-1β-induced degeneration of mouse chondrocytes. IL-1β-induced chondrocytes were transfected with pcDNA-GSK3B or together with pcDNA-NR4A3. (a) MSP-PCR assay was used to detect the methylation level of NR4A3 promoter region in chondrocytes. (b, h–j) Western blotting was used to detect the expression of NR4A3, MMP13, aggrecan, and type II collagen protein. (c) CCK-8 assay was used to detect cell viability. (d, e) Flow cytometry was used to detect apoptosis. (f, g) ELISA kits were used to detect the secretion of inflammatory factors TNF-α and IL-6. (h) RT-qPCR was used to detect the expression of MMP13, aggrecan, and type II collagen mRNA. Data is presented as the for six independent experiments and analyzed by Student’s -test or ANOVA test. and .

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